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Method for creating rice material capable of blocking rice stripe virus transmission of laodelphax striatellus

A technology of rice streak virus and striatellus planthopper, applied in the field of agricultural science, can solve the problems of insect resistance, environmental pollution, and inability to completely block virus infection and virus transmission.

Pending Publication Date: 2021-12-28
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Resistant varieties have played an important role in the control of stripe leaf blight, but resistant varieties cannot completely block the infection of plants by the virus and the transmission of the virus by the vector, and the cycle of disease infection can still be maintained in the field
Pest management can control the number of vectors and reduce the spread of viruses, but the use of chemical agents to control pests can easily cause environmental pollution and make insects resistant

Method used

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  • Method for creating rice material capable of blocking rice stripe virus transmission of laodelphax striatellus
  • Method for creating rice material capable of blocking rice stripe virus transmission of laodelphax striatellus
  • Method for creating rice material capable of blocking rice stripe virus transmission of laodelphax striatellus

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Effect test

Embodiment 1

[0013] Example 1: Using NSvc2-N S Genetically modified rice blocks transmission of RSV by SBPH

[0014] 1. NSvc2-N S gene cloning

[0015] Primers were designed according to the reported nucleotide sequence of rice stripe virus RNA2 (GenBank accession number: AY186789), NSvc2-N S -F: 5'-GGATCCATGAGGAACAGGGAGAGGGAAC-3' and NSvc2-N S -R: 5'-GGTACCTCAAGAGTACAATGAATATTT-3'. Extracting total RNA from RSV-infected rice leaves and amplifying NSvc2-N by RT-PCR S gene, and cloned the gene into the rice expression vector pBWA(V)HS to construct the transgenic expression vector pBWA(V)HS-NSvc2-N S ( figure 1 ) and verified by sequencing.

[0016] 2. Preparation of transgenic rice materials

[0017] Constructed pBWA(V)HS-NSvc2-N S The vector was transformed into Agrobacterium, and Agrobacterium was co-cultured with rice (variety: Nipponbare, Oryza sativa L.cv.Nipponbare) mature embryo callus for rice transformation, and the resistant callus was selected by hygromycin selection Tr...

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Abstract

The invention provides a method for creating a rice material capable of blocking rice stripe virus transmission of laodelphax striatellus, and belongs to the technical field of agricultural science. According to the characteristic that rice stripe virus (RSV) glycoprotein NSvc2-N-mediated virus infects mediator laodelphax striatellus midguts, NSvc2-NS segments (720bp, soluble segments encoding NSvc2-N, namely 30-268 amino acids) are selected as exogenous genes, and are integrated into rice genomes through a transgenic technology, an obtained transgenic rice plant can significantly reduce the virus obtaining rate of the laodelphax striatellus, the virus content in the laodelphax striatellus and the virus transmission rate, and proves that the NSvc2-NS protein expressed in the transgenic plant can be competitively combined with a virus infection site located in the midgut of the mediator laodelphax striatellus, so that the virus transmission of the laodelphax striatellus is blocked. The method fills up the blank of the technology of blocking the virus transmission of the laodelphax striatellus by using the plant itself, and compared with a traditional insect control and disease prevention method, the method is simple, convenient, economical, green and efficient, and can be applied to the fields of antiviral breeding work, disease prevention and control research, insect virus transmission mechanism research and the like.

Description

Technical field: [0001] The invention relates to a method of inserting rice stripe virus glycoprotein NSvc2-N into the rice genome by transgenic technology S The invention relates to a method for genetically realizing blocking the virus transmission of the small planthopper, which can be applied in agricultural production and belongs to the field of agricultural science and technology. Background technique: [0002] Rice stripe virus (RSV) belongs to the genus Tenuivirus. Its vector of transmission insects is mainly Laodelphax striatellus. . The RSV genome is composed of 4 RNAs, namely RNA1, RNA2, RNA3 and RNA4, which encode 7 proteins in total, among which the viral replicase is RdRp, NS2 and NS3 are gene silencing inhibitors, NSvc2 is glycoprotein, and NCP is the nucleocapsid Shell protein, SP is a disease-specific protein, NSvc4 is a motor protein. The main symptoms of rice infected with RSV are roughly divided into two types: rolling leaf type and leaf unfolding type....

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/40A01H5/00A01H6/46
CPCC12N15/8286C07K14/005C12N2760/00022C12N2760/00031
Inventor 李硕卢成晔季英华周益军
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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